The purpose of the current research was to study the influence of dietary supplementation of nettle (Urtica dioica) on laying performance, egg quality and blood serum biochemical parameters of layers. A total of 60 laying hens (42 weeks old) from Lohman Klassik Brown breed were randomly allocated into three groups: a control and two experimental groups (n = 20 hens per group). All layers received compound feed with the following nutritional value: 2710 Kcal/kg metabolizable energy; 16.44% crude protein; 3.32% crude fats; 4.58% crude fibres; 3.73% Са; 0.49% Р. The hens from the experimental groups received 0.3% (experimental group 1) and 0.5% (experimental group 2) of dried nettle with the diet. Both experimental groups had significantly higher egg yolk pigmentation (p < 0.001) compared with the control group. A significantly lower egg yolk cholesterol content was found in hens from experimental group 1 (p < 0.05). Nettle addition reduced significantly blood serum glucose (p < 0.01 and p < 0.05 in experimental groups 1 and 2, respectively) as well as the total serum cholesterol content (p < 0.001).